Friedman P N, Chen X, Bargonetti J, Prives C
Department of Biological Sciences, Columbia University, New York, NY 10027.
Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3319-23. doi: 10.1073/pnas.90.8.3319.
We have analyzed the size and structure of native immunopurified human p53 protein. By using a combination of chemical crosslinking, gel filtration chromatography, and zonal velocity gradient centrifugation, we have determined that the predominant form of p53 in such preparations is a tetramer. The behavior of purified p53 in gels and sucrose gradients implies that the protein has an extended shape. Wild-type p53 has been shown to bind specifically to sites in cellular and viral DNA. We show in this study by Southwestern ligand blotting and by analysis of DNA-bound crosslinked p53 that p53 monomers, dimers, and tetramers can bind directly to DNA.
我们分析了天然免疫纯化的人p53蛋白的大小和结构。通过化学交联、凝胶过滤色谱和区带速度梯度离心相结合的方法，我们确定了此类制剂中p53的主要形式是四聚体。纯化的p53在凝胶和蔗糖梯度中的行为表明该蛋白具有伸展的形状。野生型p53已被证明能特异性结合细胞和病毒DNA中的位点。我们在本研究中通过蛋白质印迹法和对与DNA结合的交联p53的分析表明，p53单体、二聚体和四聚体均可直接结合DNA。
